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Molecular Genetic Studies of Thrombosis

血栓形成的分子遗传学研究

基本信息

批准号：
01480298
负责人：
KOIDE Takehiko
金额：
$ 4.35万
依托单位：
Himeji Institute of Technology (1991)Niigata University (1989-1990)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480298/
关键词：
Thrombosis Protein C Protein C deficiency Polymorphism Gene analysis Histidine-rich glycoprotein Protein C inhibitor Chromosome 3 プロテインC異常症 HRG PCR PCR法

项目摘要

1. Analysis of Genetic Abnormalities for Protein C Deficiency. Makinal use of the polymerase chain reaction (PCR), we analyzed the genes for protein C from Japanese patients with a hereditary protein C deficiency, and identified two molecular abnormalities in two independent probands. One is a 3414T to C mutation in exon VI which converts Tyrl24 (ZAC) to His (CAC) in the second EGF domain. The other is a frameshift deletion of a single G nucleotide in a region of 8854G-8857G in exon IX coding for the carboxy-terminal region, which is predicted to result in an alteration and elongation of the deduced carboxy-terminal amino acid sequence.2. Diagnosis of Familial Protein C Deficiency. Based on a result described above, we have established a direct diagnostic method of familial protein C deficiency in the second case of a frameshift deletion by use of mutacenic primers for PCR to introduce Ava I or Bal I site into the amplified product.3. Polymorphism in the Protein C Gene. We have identified two sequence variations in the protein C gene. The first one is an A-T sequence polymorphism at nucleotide -1476 in the untranslated exon 1, and the second one is a T-G sequence polymorphism in the triplet coding for Ser-99 (TCT - TCG).4. Heparin-Binding Property of Protein C. We found that protein C, and more strongly activated protein C, interacts with heparin, which is enhanced in the presence of calcium ions, and proposed a specific region of protein C as a heparin-binding site.5. Modulation of Protein C Inhibitor Activity by Histidine-Rich Glycoprotein (HRG). We have found that HRG neutralizes the heparin-dependent inhibitions of activated protein C and thrombin by protein C inhibitor at physioloical concentrations of zinc and calcium ions in plasma, suggesting a new function of HRG as a physiological modulator of protein C inhibitor.6. Assignment of the Gene for HRG to Chromosome 3. We have assigned the gene for HRG to chromosome 3.

1.蛋白C缺乏症的遗传异常分析最大限度地使用聚合酶链反应（PCR），我们分析了日本遗传性蛋白C缺乏症患者的蛋白C基因，并确定了两个独立的先证者的两个分子异常。一个是外显子VI中的3414 T至C突变，其将第二EGF结构域中的Tyr 124（ZAC）转化为His（CAC）。另一种是外显子IX中编码羧基端的8854 G-8857 G区域的一个G核苷酸移码缺失，预计会导致推导的羧基端氨基酸序列的改变和延长.家族性蛋白C缺乏症的诊断基于上述结果，我们建立了一种直接诊断第二例移码缺失的家族性蛋白C缺乏症的方法，即使用突变引物进行PCR，将Ava I或Bal I位点引入扩增产物中.蛋白C基因的多态性我们已经确定了蛋白C基因中的两个序列变异。第一个是非翻译外显子1中-1476位核苷酸处的A-T序列多态性，第二个是编码Ser-99的三联体中的T-G序列多态性（TCT-TCG）.蛋白C的肝素结合特性。我们发现蛋白C和更强活化的蛋白C与肝素相互作用，这在钙离子存在下增强，并提出蛋白C的特定区域作为肝素结合位点.富含组氨酸的糖蛋白（HRG）对蛋白C抑制剂活性的调节。我们发现HRG在生理浓度的锌离子和钙离子存在下能中和蛋白C抑制剂对活化蛋白C和凝血酶的肝素依赖性抑制，提示HRG作为蛋白C抑制剂的一种新的生理调节剂.将HRG基因分配至3号染色体。我们已将HRG基因定位于3号染色体。